全文获取类型
收费全文 | 579篇 |
免费 | 20篇 |
出版年
2024年 | 3篇 |
2022年 | 5篇 |
2021年 | 4篇 |
2020年 | 6篇 |
2019年 | 6篇 |
2018年 | 8篇 |
2016年 | 11篇 |
2015年 | 9篇 |
2014年 | 29篇 |
2013年 | 31篇 |
2012年 | 33篇 |
2011年 | 28篇 |
2010年 | 15篇 |
2009年 | 36篇 |
2008年 | 35篇 |
2007年 | 38篇 |
2006年 | 33篇 |
2005年 | 29篇 |
2004年 | 30篇 |
2003年 | 25篇 |
2002年 | 22篇 |
2001年 | 18篇 |
2000年 | 17篇 |
1999年 | 11篇 |
1998年 | 10篇 |
1997年 | 9篇 |
1996年 | 7篇 |
1995年 | 4篇 |
1993年 | 3篇 |
1992年 | 7篇 |
1991年 | 10篇 |
1990年 | 4篇 |
1989年 | 3篇 |
1988年 | 8篇 |
1987年 | 4篇 |
1986年 | 3篇 |
1985年 | 2篇 |
1984年 | 4篇 |
1983年 | 4篇 |
1982年 | 4篇 |
1978年 | 3篇 |
1977年 | 4篇 |
1973年 | 4篇 |
1971年 | 4篇 |
1970年 | 2篇 |
1969年 | 2篇 |
1968年 | 2篇 |
1966年 | 1篇 |
1965年 | 2篇 |
1961年 | 1篇 |
排序方式: 共有599条查询结果,搜索用时 31 毫秒
101.
Chikara Kubota Xiangzhong Yang Andras Dinnyes Junichi Todoroki Hiroshi Yamakuchi Kazunori Mizoshita Satoru Inohae Norio Tabara 《Molecular reproduction and development》1998,51(3):281-286
Cryopreservation of bovine oocytes would be beneficial both for nuclear transfer and for preservation efforts. The overall objective of this study was to evaluate the viability as well as the cryodamage to the nucleus vs. cytoplasm of bovine oocytes following freezing-thawing of oocytes at immature (GV) and matured (MII) stages using in vitro fertilization (IVF), parthenogenetic activation, or nuclear transfer assays. Oocytes were collected from slaughterhouse ovaries. Oocytes at the GV, MII, or MII but enucleated (MIIe) stages were cryopreserved in 5% (v/v) ethylene glycol; 6% (v/v) 1,2-propanediol; and 0.1-M sucrose in PBS supplemented with 20% (v/v) fetal bovine serum. Frozen-thawed oocytes were subjected to IVF, parthenogenetic activation, or nuclear transfer assays. Significantly fewer GV oocytes survived (i.e., remained morphologically intact during freezing-thawing) than did MII oocytes (47% vs. 84%). Subsequent development of the surviving frozen-thawed GV and MII oocytes was not different (58% and 60% cleavage development; 7% and 12% blastocyst development at Day 9, respectively, P > 0.05). Parthenogenetic activation of frozen-thawed oocytes resulted in significantly lower rates of blastocyst development for the GV than the MII oocyte groups (1% vs. 14%). Nuclear transfer with cytoplasts derived from frozen-thawed GV, MII, MIIe, and fresh-MII control oocytes resulted in 5%, 16%, 14%, and 17% blastocyst development, respectively. However, results of preliminary embryo transfer trials showed that fewer pregnancies were produced from cloned embryos derived from frozen oocytes or cytoplasts (9%, n = 11 embryos) than from fresh ones (19%, n = 21 embryos). Transfer of embryos derived by IVF from cryopreserved GV and MII oocytes also resulted in term development of calves. Our results showed that both GV and MII oocytes could survive freezing and were capable of developing into offspring following IVF or nuclear transfer. However, blastocyst development of frozen-thawed oocytes remains poorer than that of fresh oocytes, and our nuclear transfer assay suggests that this poorer development was likely caused by cryodamage to the oocyte cytoplasm as well as to the nucleus. Mol. Reprod. Dev. 51:281–286, 1998. © 1998 Wiley-Liss, Inc. 相似文献
102.
Noriaki Sasai Shogo Tada Jumi Ohshiro Chikara Kogiso Takuma Shinozuka 《Development, growth & differentiation》2024,66(1):89-100
During development, progenitor cell survival is essential for proper tissue functions, but the underlying mechanisms are not fully understood. Here we show that ERCC6L2, a member of the Snf2 family of helicase-like proteins, plays an essential role in the survival of developing chick neural cells. ERCC6L2 expression is induced by the Sonic Hedgehog (Shh) signaling molecule by a mechanism similar to that of the known Shh target genes Ptch1 and Gli1. ERCC6L2 blocks programmed cell death induced by Shh inhibition and this inhibition is independent of neural tube patterning. ERCC6L2 knockdown by siRNA resulted in the aberrant appearance of apoptotic cells. Furthermore, ERCC6L2 cooperates with the Shh signal and plays an essential role in the induction of the anti-apoptotic factor Bcl-2. Taken together, ERCC6L2 acts as a key factor in ensuring the survival of neural progenitor cells. 相似文献
103.
Prolactin induces MFG-E8 production in macrophages via transcription factor C/EBPβ-dependent pathway
Aziz MM Ishihara S Rumi MA Mishima Y Oshima N Kadota C Moriyama I Li YY Rahman FB Otani A Oka A Ishimura N Kadowaki Y Amano Y Kinoshita Y 《Apoptosis : an international journal on programmed cell death》2008,13(5):609-620
The lactogenic hormone prolactin (PRL) regulates milk protein gene expression in mammary glands. To maintain homeostatic balance
in the body, milk fat globule epidermal growth factor 8 (MFG-E8) is vital for phagocytic clearance of apoptotic cells. We
investigated the effects of PRL on MFG-E8 expression in macrophages by evaluating its promoter function. Macrophages were
stimulated with PRL, and the expression of MFG-E8 was determined using real-time PCR and Western blotting. The role of MFG-E8
on phagocytosis of apoptotic cells in PRL-treated macrophages was assessed using microscopy, while the response of PRL to
MFG-E8 expression was evaluated using luciferase assay. Following treatment with PRL, significant up-regulations of the PRL
receptor and MFG-E8 were observed in macrophages, though PRL-treated macrophages more efficiently engulfed apoptotic cells.
The results of MFG-E8 promoter analysis showed considerable up-regulation of promoter activity in macrophages following PRL
treatment and results from mutation analysis of the MFG-E8 promoter suggested that the C/EBPβ binding site was responsible
for PRL-induced activation of the MFG-E8 promoter. C/EBPβ activity was found to be up-regulated in PRL-treated cells as revealed
by an electrophoretic mobility shift assay (EMSA). In conclusion, PRL is a potent inducer of MFG-E8 expression in macrophages,
while its effect is mediated by the presence of a responsive element in the MFG-E8 promoter. 相似文献
104.
We propose a principle of consistency between different hierarchical levels of biological systems. Given a consistency between
molecule replication and cell reproduction, universal statistical laws on cellular chemical abundances are derived and confirmed
experimentally. They include a power law distribution of gene expressions, a lognormal distribution of cellular chemical abundances
over cells, and embedding of the power law into the network connectivity distribution. Second, given a consistency between
genotype and phenotype, a general relationship between phenotype fluctuations by genetic variation and isogenic phenotypic
fluctuation by developmental noise is derived. Third, we discuss the chaos mechanism for stem cell differentiation with autonomous
regulation, resulting from a consistency between cell reproduction and growth of the cell ensemble.
相似文献
Kunihiko KanekoEmail: |
105.
How can a microorganism adapt to a variety of environmental conditions despite the existence of a limited number of signal transduction mechanisms? We show that for any growing cells whose gene expression fluctuate stochastically, the adaptive cellular state is inevitably selected by noise, even without a specific signal transduction network for it. In general, changes in protein concentration in a cell are given by its synthesis minus dilution and degradation, both of which are proportional to the rate of cell growth. In an adaptive state with a higher growth speed, both terms are large and balanced. Under the presence of noise in gene expression, the adaptive state is less affected by stochasticity since both the synthesis and dilution terms are large, while for a nonadaptive state both the terms are smaller so that cells are easily kicked out of the original state by noise. Hence, escape time from a cellular state and the cellular growth rate are negatively correlated. This leads to a selection of adaptive states with higher growth rates, and model simulations confirm this selection to take place in general. The results suggest a general form of adaptation that has never been brought to light—a process that requires no specific mechanisms for sensory adaptation. The present scheme may help explain a wide range of cellular adaptive responses including the metabolic flux optimization for maximal cell growth. 相似文献
106.
Yamaoka M Uematsu T Shiba T Matsuura T Ono Y Ishizuka M Naramoto H Takahashi M Sugiura-Tomita M Iguchi K Yamashita S Furusawa K 《Gerodontology》2008,25(1):10-17
Aim: Inorganic polyphosphate exists as chains of phosphate molecules and is distributed in osteoblasts, and regulates osteoblastic cell differentiation and bone matrix calcification. The purpose of this study was to clarify the effects of inorganic polyphosphate on periodontitis. Material and methods: Subgingival local irrigation with inorganic polyphosphate was studied in a randomised double‐blind study of 33 patients with periodontitis. Scaling and root planing were performed 1 week after the initial examination. Results: No significant differences between the inorganic polyphosphate group and control were detected in each item except IL‐1β. Patients in whom both the bleeding on probing and gingival index at 1 week had improved were significantly older in the inorganic polyphosphate group than in the control group (p < 0.05). Bone regeneration was seen in one case of the inorganic polyphosphate group. Conclusions: Inorganic polyphosphate was useful in the treatment of periodontitis in the elderly, indicating a probable effect of anti‐ageing, with similar bone regenerations occurring in both groups. 相似文献
107.
108.
Zhipeng Wang Yuhei Nishimura Yasuhito Shimada Noriko Umemoto Minoru Hirano Liqing Zang Takehiko Oka Chikara Sakamoto Junya Kuroyanagi Toshio Tanaka 《Gene》2009
Beta adrenergic receptors (β-ARs) are members of the G-protein-coupled receptor superfamily and mediate various physiological processes in many species. The expression patterns and functions of β-ARs in zebrafish are, however, largely unknown. We have identified zebrafish β-AR orthologs, which we have designated as adrb1, adrb2a, adrb2b, adrb3a and adrb3b. adrb1 was found to be expressed in the heart and brain. Expression of adrb2a predominated in the brain and skin, whereas adrb2b was found to be highly expressed in muscle, pancreas and liver. Both adrb3a and adrb3b were exclusively expressed in blood. Knock-down of these β-ARs by morpholino oligonucleotides revealed a functional importance of adrb2a in pigmentation. Expression of atp5a1 and atp5b, genes that encode subunits of F1F0-ATPase, which is known to be involved in pigmentation, was significantly increased by knock-down of adrb2a. Our data suggest that adrb2a may regulate pigmentation, partly by modulating F1F0-ATPase. 相似文献
109.
Shin Iwamoto Norihisa Nishimichi Yoshiko Tateishi Yuko Sato Hiroyuki Horiuchi Shuichi Furusawa Tatsuya Sawamura Haruo Matsuda 《MABS-AUSTIN》2009,1(4):357-363
Lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (LOX-1) is the major receptor for oxidized LDL (oxLDL), and plays a key role in the pathogenesis of atherosclerosis and cardiovascular diseases. Monoclonal antibodies (mAbs) specific for human LOX-1 (hLOX-1) were generated by a phage display technique using chickens immunized with recombinant hLOX-1 (rhLOX-1). A total of 53 independent scFv clones reactive for rhLOX-1 were obtained. Of the 53 clones, 49 recognized the C-type lectin-like domain (CTL domain), which contributes to the binding of oxLDL. Of these, 45 clones inhibited oxLDL-binding with LOX-1. Furthermore, some of these clones cross-reacted with rabbit, pig and/or mouse LOX-1. For possible application as therapeutic agents in the future, two cross-reactive mAbs were re-constructed as chicken-human chimeric antibodies. The chimeric antibodies showed similar characteristics compared to the original antibodies, and inhibited oxLDL binding to LOX-1 expressed on CHO cells. The results obtained in this study indicate that anti-LOX-1 mAbs might be useful tools for functional analyses and development of therapeutic agents for cardiovascular indications such as atherosclerosis.Key words: LOX-1, oxLDL, chicken monoclonal antibody, chimeric antibody, neutralizing antibody 相似文献
110.
Yuusuke Maruyama Toshihiko Ogura Kazuhiro Mio Kenta Kato Takeshi Kaneko Shigeki Kiyonaka Yasuo Mori Chikara Sato 《The Journal of biological chemistry》2009,284(20):13676-13685
The Ca2+ release-activated Ca2+ channel is a
principal regulator of intracellular Ca2+ rise, which conducts
various biological functions, including immune responses. This channel,
involved in store-operated Ca2+ influx, is believed to be composed
of at least two major components. Orai1 has a putative channel pore and
locates in the plasma membrane, and STIM1 is a sensor for luminal
Ca2+ store depletion in the endoplasmic reticulum membrane. Here we
have purified the FLAG-fused Orai1 protein, determined its tetrameric
stoichiometry, and reconstructed its three-dimensional structure at 21-Å
resolution from 3681 automatically selected particle images, taken with an
electron microscope. This first structural depiction of a member of the Orai
family shows an elongated teardrop-shape 150Å in height and 95Å in
width. Antibody decoration and volume estimation from the amino acid sequence
indicate that the widest transmembrane domain is located between the round
extracellular domain and the tapered cytoplasmic domain. The cytoplasmic
length of 100Å is sufficient for direct association with STIM1. Orifices
close to the extracellular and intracellular membrane surfaces of Orai1 seem
to connect outside the molecule to large internal cavities.Ca2+ is an intracellular second messenger that plays important
roles in various physiological functions such as immune response, muscle
contraction, neurotransmitter release, and cell proliferation. Intracellular
Ca2+ is mainly stored in the endoplasmic reticulum
(ER).2 This ER system
is distributed through the cytoplasm from around the nucleus to the cell
periphery close to the plasma membrane. In non-excitable cells, the ER
releases Ca2+ through the inositol 1,4,5-trisphosphate
(IP3) receptor channel in response to various signals, and the
Ca2+ store is depleted. Depletion of Ca2+ then induces
Ca2+ influx from outside the cell to help in refilling the
Ca2+ stores and to continue Ca2+ rise for several
minutes in the cytoplasm (1,
2). This Ca2+ influx
was first proposed by Putney
(3) and was named
store-operated Ca2+ influx. In the immune system, store-operated
Ca2+ influx is mainly mediated by the Ca2+
release-activated Ca2+ (CRAC) current, which is a highly
Ca2+-selective inwardly rectified current with low conductance
(4,
5). Pathologically, the loss of
CRAC current in T cells causes severe combined immunodeficiency
(6) where many Ca2+
signal-dependent gene expressions, including cytokines, are interrupted
(7). Therefore, CRAC current is
necessary for T cell functions.Recently, Orai1 (also called CRACM1) and STIM1 have been physiologically
characterized as essential components of the CRAC channel
(8–12).
They are separately located in the plasma membrane and in the ER membrane;
co-expression of these proteins presents heterologous CRAC-like currents in
various types of cells (10,
13–15).
Both of them are shown to be expressed ubiquitously in various tissues
(16–18).
STIM1 senses Ca2+ depletion in the ER through its EF hand motif
(19) and transmits a signal to
Orai1 in the plasma membrane. Although Orai1 is proposed as a regulatory
component for some transient receptor potential canonical channels
(20,
21), it is believed from the
mutation analyses to be the pore-forming subunit of the CRAC channel
(8,
22–24).
In the steady state, both Orai1 and STIM1 molecules are dispersed in each
membrane. When store depletion occurs, STIM1 proteins gather into clusters to
form puncta in the ER membrane near the plasma membrane
(11,
19). These clusters then
trigger the clustering of Orai1 in the plasma membrane sites opposite the
puncta (25,
26), and CRAC channels are
activated (27).Orai1 has two homologous genes, Orai2 and Orai3
(8). They form the Orai family
and have in common the four transmembrane (TM) segments with relatively large
N and C termini. These termini are demonstrated to be in the cytoplasm,
because both N- and C-terminally introduced tags are immunologically detected
only in the membrane-permeabilized cells
(8,
9). The subunit stoichiometry
of Orai1 is as yet controversial: it is believed to be an oligomer, presumably
a dimer or tetramer even in the steady state
(16,
28–30).Despite the accumulation of biochemical and electrophysiological data,
structural information about Orai1 is limited due to difficulties in
purification and crystallization. In this study, we have purified Orai1 in its
tetrameric form and have reconstructed the three-dimensional structure from
negatively stained electron microscopic (EM) images. 相似文献